Prevention of Osteoarthritis (OA)
From a human perspective, OA is a crippling disease with high socio-economic impact. OA is a heterogeneous group of conditions, including Primary OA (e.g. genitive factors, hormonal factors, mechanical stress to joints) and Secondary OA (e.g. inflammatory episodes, post-traumatic).
It is the leading cause of disability with about 43 million people affected in the US and 240 million worldwide. It has substantial economic impact, e.g. in the US direct treatment costs $ 10.7 billion, indirect costs $ 42.8 billion (missed working days). Symptoms are pain, hypertrophy and stiffness of joints and limitation of movement. Current therapy focuses on pain relief with non-steroidal anti-inflammatory drugs (NSAIDs).
From a pet perspective, osteoarthritis is an age- and weight-related degenerative problem of the joints. It affects 20 Mio dogs worldwide: dogs have trouble in getting up and jumping, are sore after exercise, and/or grumpy. Because of joint degeneration, inflammation sets on.
Natural therapeutic treatments (glucosamine and glycosaminoglycans) are effective in slowing progression of disease.
Use of Glucosamine
The use of pure glucosamine in the treatment of joint diseases is widely described in the patent as well as in the scientific literature, usually in combination with other compounds or extracts from various natural sources. Pure glucosamine is added as glucosamine hydrochloride or glucosamine sulphate, and comes from shellfish hydrolysis. For example, WO20000074696 describes “herbal compositions comprising glucosamine and Trypterygium wilfordii, Ligustrum lucidum and/or Erycibe schmidtii, for treating inflammation or degeneration of joint tissues, e.g. arthritis” where pure glucosamine is mixed with plant preparation. Other patents relate to compositions of plant carbohydrates as dietary supplements (EP1172041 or EP0923382) where glucosamine is originated from chitin.
The use of glucosamine as an anti-osteoarthritis agent has been intensively developed during the last decade. Glucosamine is suspected to be one of the sole active compound on joint disease such as osteoarthritis (up to recently only symptomatic treatment such as non-steroidal anti-inflammatory drugs have been sought to be efficient).
Glucosamine has also been shown preventing the cartilage degradation by inhibiting the production of MMPs (Matrix metalloproteases) such as MMP1, MMP3 and MMP13
Interestingly glucosamine is also related to the aging process of skin, which has been characterized mainly by the continuous loss of elasticity and the loss of moisture. Skin aging is reflected by major structural changes and variations in composition. Most notably aged skin has less collagen and glycosaminoglycans compared with young skin. Glycosaminoglycan molecules produced by the skin include hyaluronic acid (poly d-glucuronic acid-n-acetyl-d-glucosamine), chondroitin sulfate, and dermatan sulfate. Hyaluronic acid is produced in higher quantities by the skin cells in response to exfoliation. Hyaluronic acid has a large capacity for hydration.
Inhibiting MMP-1 is related to the inhibition of the polyglycan/collagen degradation, and therefore also related to skin ageing: MMP-1 can be induced by UV and is recognized as a marker of the skin ageing. In US2002119107, an invention is based on the selective inhibition of MMP-1 claiming topical compositions for protecting human skin from collagen degradation. US2004037901 claims a regime for inhibiting the adverse signs of effects of cutaneous aging comprising an extract from rosemary plant inhibiting the expression of metalloproteases.
Glucosamine has been shown to significantly improve dryness of the skin and exfoliation. Glucosamine increases the moisture content and improves the smoothness of the skin. These findings suggest that long-term intake of glucosamine is effective in improving moisture content and smoothness of the skin.
It has been shown that oral supplement containing glucosamine lead to a reduction (34%) in the number of visible wrinkles and (34%) in the number of fine lines in a group of women who took the supplement. The use of an oral supplement containing glucosamine, minerals, and various antioxidant compounds can potentially improve the appearance of visible wrinkles and fine lines.
U.S. Pat. No. 6,413,525, describes methods of substantially exfoliating the skin. In particular, the invention relates to topically applied compositions containing an amino sugar in the form of N-acetyl glucosamine: when young skin cells are exposed after exfoliation, they produce larger quantities of hyaluronic acid which is a glycosaminoglycan composed of a chain of alternating, repeating, D-glucuronic acid and N-acetyl-D-glucosamine molecules. N-acetyl-D-glucosamine is known to be a rate-limiting factor in the hyaluronic acid production by living cells. The topical application of glucosamine assists in the continued production of hyaluronic acid.
Other Compositions for topical application containing N-acetyl-D-glucosamine have been also disclosed for example, in JP 59013708 (soften and moisturize the skin) U.S. Pat. No. 5,866,142 (a composition for exfoliating the skin).
Origin of Glucosamine
Glucosamine is an amino monosaccharide found in chitin, glycoproteins, proteoglycans (PGs), and glycosaminoglycans (GAGs) such as hyaluronic acid. Glucosamine is also known as 2-amino-2-deoxyglucose, 2-amino-2-deoxy-beta-D-glucopyranose and chitosamine. It has the following formula:

Glucosamine in Animals:
Glucosamine is a constituent of glycosaminoglycans in cartilage matrix and synovial fluids. They are in form of polymers of glucosamine, with an acetyl group attached to a variable number of the individual glucosamine molecules (making them acetylglucosamine).
A polymer composed totally of acetylglucosamine is called chitin, and one composed totally of glucosamine is called chitosan.
The structure of chitin is very similar to cellulose, especially because they both are β (1-4) linked.
The main sources of chitin are the following (in % of total dry matter)
Fungi5-20%Spiders38%Worms20-38% Cockroaches35%Squids/Octopus3-20%Water Beetle37%Scorpions30%Silk Worm44%Edible Crab70%Hermit Crab69%
Glucosamine in Plants:
N-Glycosylproteins, which are N-linked glycoproteins, are present in plants in trace amounts. For example, a glucosamine residue of oligosaccharide is N-glycosidically attached to the amide nitrogen of an asparagine residue of the protein. Examples are phaseolin, legumin, bromelain, laccase, etc. Degradation occurs through the activity of enzymes for de-N-glycosylation (cleavage of glucosamine linkage between N-acetyl-D glucosamine and Asp. residue).
N-Glycosylproteins are different from chitin, found in the extracellular matrix, in the vacuole, associated to membranes (Endoplasmic Reticulum, Golgi, tonoplast, plasma membrane). N-glycans influence the stability, solubility, and biological activity of the protein. De-N-glycosylation seems important during germination and post germinative development.
Such a linked-glucosamine is in limited quantities and not freely available (or through hydrolysis such as strong acidic or enzymatic treatment).
Free glucosamine has not been observed in plant species or only as trace levels: (for examples less than 1 mg/kg dry matter in fresh chicory, carrot, Jerusalem artichoke or beet raw materials, or commercial dried chicory roots such as those from Leroux company (France). The publication of Alabran D. M. and Mabrouk A. F. (Carrot flavor. Sugars and free nitrogenous compounds in fresh carrots, J. Agr. Food Chem., 21 (2), 205-208, 1973) is the only scientific publication describing the presence of free glucosamine in fresh (non-processed, non-dried material) plant material in a relatively high amount (0.071% of fresh carrot). Nevertheless, surprisingly there is no other data reporting free glucosamine on carrot or other plant material in the scientific literature or referenced Nutrition Tables such as “Food Composition and Nutrition Tables, MedPharm (Stuttgart) and CRC Press (Boca Raton), 1994”. Furthermore, our own investigation detected less than 1 mg/kg dry matter in commercial root of carrots, and confirm the state of the art in that glucosamine is only present at a level of traces in vegetables. In WO2003/070168, carrot seeds have been studied for their chemical composition and their activity for the reduction of inflammation-related pain, without any reference to glucosamine. It does not concern a food or pet food composition containing glucosamine generated from plant materials through a drying process as set forth in our present invention.
Chicory is used in several food/pet food compositions, using commercial dried, comminuted roots, and usually does not contain significant amount of glucosamine. For example, EP 0850569A1 describes a gelatinised cereal product containing oligosaccharide using chicory root as one of the ingredients. Chicory is used as a source of inulin and fructooligosaccharides and in use the cereal product has a beneficial effect in the gastrointestinal tracts of human and animals. In US2004/0001898A, food compositions, containing chicory, are described for detoxification and cancer prevention.
In EP1325682A1, various plant extracts are used in food or pet food compositions for maintenance of bone health. As in many other food/pet food compositions, using commercial dried, comminuted roots, not containing glucosamine, Chicory is cited only as one of the ingredients of a pet food composition (example 3), not as the ingredient responsible of the claimed biological activity and without any reference to generation of glucosamine or presence or use of glucosamine in plant material or final product or to a specific drying process.
In JP63309147, chicory material is hydrolysed with an acid to generate oligofructosaccharides. It does not refer to glucosamine generation or presence or use of glucosamine in plant material or final product.
In HU66929, a natural fodder additive is described, including oats-flour, apple juice, dried bean pods and comminuted dried Jerusalem artichoke. The claimed use is for lowering cholesterol levels in animal products. It does not refer to glucosamine generation or presence or use of glucosamine in plant material or final product.
In WO2003/070168, carrot seeds have been studied for their chemical composition and their activity for the reduction of inflammation-related pain (cycloxygenase enzyme-mediated inflammation). It does not refer to glucosamine generation or presence or use of glucosamine in carrot raw material or extract or isolated compounds.
Industrial Sources of Glucosamine
Industrial glucosamine is a pure compound obtained from the acidic hydrolysis of chitin from shellfish, a complex carbohydrate derived from N-acetyl-D-glucosamine. Glucosamine can also be produced from enzymatic hydrolysis of shellfish, microbial fermentation (for example with of corn-derived products).
Patents have been filed protecting fermentation processes (thus micro organisms) leading to the production of glucosamine. All these processes concern the production of pure, extracted glucosamine, in competition with shellfish extracts.
As an example, U.S. Pat. No. 6,486,307 describes an improved method for chitin acidic hydrolysis: A method of producing glucosamine hydrochloride from chitin by grinding the chitin to a very fine size and digestion with concentrated hydrochloric acid.
U.S. Pat. No. 6,372,457 describes a method and material for producing glucosamine by fermentation of a genetically modified microorganism.
U.S. Pat. No. 5,998,173 describes a novel process for directly producing N-acetyl-D-glucosamine from chitin utilizing an ensemble of the chitinase family of enzymes to hydrolyse chitin of crustacean shells.